CA3099508A1 - Procede de realisation d'analyse de cellules uniques et dispositif associe - Google Patents

Procede de realisation d'analyse de cellules uniques et dispositif associe Download PDF

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Publication number
CA3099508A1
CA3099508A1 CA3099508A CA3099508A CA3099508A1 CA 3099508 A1 CA3099508 A1 CA 3099508A1 CA 3099508 A CA3099508 A CA 3099508A CA 3099508 A CA3099508 A CA 3099508A CA 3099508 A1 CA3099508 A1 CA 3099508A1
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Prior art keywords
cell
gel capsule
cells
gel
genome
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CA3099508A
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Masahito Hosokawa
Haruko Takeyama
Yohei Nishikawa
Masato Kogawa
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BitBiome Inc
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BitBiome Inc
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Publication of CA3099508A1 publication Critical patent/CA3099508A1/fr
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1075—Isolating an individual clone by screening libraries by coupling phenotype to genotype, not provided for in other groups of this subclass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
    • B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
    • B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B01L3/502792—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics for moving individual droplets on a plate, e.g. by locally altering surface tension
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    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C—CHEMISTRY; METALLURGY
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    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0652—Sorting or classification of particles or molecules
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0663—Stretching or orienting elongated molecules or particles
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/069—Absorbents; Gels to retain a fluid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2547/00—Reactions characterised by the features used to prevent contamination
    • C12Q2547/10—Reactions characterised by the features used to prevent contamination the purpose being preventing contamination
    • C12Q2547/101—Reactions characterised by the features used to prevent contamination the purpose being preventing contamination by confinement to a single tube/container

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Dispersion Chemistry (AREA)
  • Immunology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Plant Pathology (AREA)
  • Clinical Laboratory Science (AREA)
  • Hematology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un procédé de production de bibliothèque de génome dans lequel une lyse cellulaire et une amplification du génome sont effectuées à l'aide d'une opération simple. Plus particulièrement, la présente invention concerne un procédé qui est destiné à amplifier des polynucléotides dans des cellules et qui comprend : une étape consistant à utiliser un échantillon contenant deux cellules ou plus ou des structures de type cellule, et encapsuler les cellules ou les structures de type cellule dans des gouttelettes, une pour chaque gouttelette ; une étape pour gélifier les gouttelettes pour générer des capsules de gel ; une étape pour effectuer une lyse des cellules ou des structures de type cellule par immersion des capsules de gel dans un ou plusieurs types de réactifs pour une lyse de façon à amener les polynucléotides dans les cellules à être élués dans les capsules de gel et à être maintenus dans les capsules de gel dans un état dans lequel des substances se liant aux polynucléotides sont éliminées ; et une étape pour amener les polynucléotides en contact avec un réactif pour une amplification à amplifier les polynucléotides dans les capsules de gel.
CA3099508A 2018-05-07 2019-04-26 Procede de realisation d'analyse de cellules uniques et dispositif associe Pending CA3099508A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
JP2018-089259 2018-05-07
JP2018089259 2018-05-07
PCT/JP2019/017952 WO2019216271A1 (fr) 2018-05-07 2019-04-26 Procédé de réalisation d'analyse de cellules uniques et dispositif associé

Publications (1)

Publication Number Publication Date
CA3099508A1 true CA3099508A1 (fr) 2019-11-14

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CA3099508A Pending CA3099508A1 (fr) 2018-05-07 2019-04-26 Procede de realisation d'analyse de cellules uniques et dispositif associe

Country Status (10)

Country Link
US (1) US11142787B2 (fr)
EP (1) EP3812460A4 (fr)
JP (2) JP6877804B2 (fr)
KR (1) KR20210006418A (fr)
CN (1) CN112334578A (fr)
AU (1) AU2019265111A1 (fr)
CA (1) CA3099508A1 (fr)
SG (1) SG11202011073RA (fr)
TW (1) TW202003829A (fr)
WO (1) WO2019216271A1 (fr)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPWO2020218551A1 (fr) * 2019-04-26 2020-10-29
JPWO2020218554A1 (fr) * 2019-04-26 2020-10-29
WO2020218553A1 (fr) * 2019-04-26 2020-10-29 ｂｉｔＢｉｏｍｅ株式会社 Analyse numérique de la flore microbienne
WO2021128052A1 (fr) * 2019-12-25 2021-07-01 苏州绘真生物科技有限公司 Kit pour construire une banque de séquençage bcr de cellules isolées humaines et ses applications
JP7460203B2 (ja) * 2020-11-11 2024-04-02 国立研究開発法人物質・材料研究機構 細胞で産生された膜小胞に由来する塩基配列を解析する方法、その装置、及び、そのプログラム
JP7473251B2 (ja) 2020-12-16 2024-04-23 国立研究開発法人物質・材料研究機構 ゲノム解析方法
KR102529008B1 (ko) * 2021-03-30 2023-05-08 숙명여자대학교산학협력단 핵산 추출용 조성물 및 이를 이용한 핵산 추출 방법
CN114441264B (zh) * 2022-01-20 2023-05-30 复旦大学 一种皮升级体积单细胞样品裂解酶解反应器
WO2023183327A1 (fr) * 2022-03-22 2023-09-28 Cellanome, Inc. Dosages cellulaires multiplex pour le criblage et l'évaluation de qualité de cellules modifiées
WO2024075787A1 (fr) * 2022-10-04 2024-04-11 ｂｉｔＢｉｏｍｅ株式会社 Procédé hautement efficace pour obtenir une séquence génique

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2504448B1 (fr) 2009-11-25 2016-10-19 Bio-Rad Laboratories, Inc. Procédés et compositions destinés à détecter un matériel génétique
WO2015069798A1 (fr) 2013-11-05 2015-05-14 The Regents Of The University Of California Typage médico-légal par microsatellites d'une cellule isolée présente dans des gouttelettes microfluidiques
JP2017532024A (ja) 2014-09-09 2017-11-02 ザ・ブロード・インスティテュート・インコーポレイテッド コンポジット単一細胞核酸分析のための液滴ベースの方法および機器
KR20200020997A (ko) * 2015-02-10 2020-02-26 일루미나, 인코포레이티드 세포 성분을 분석하기 위한 방법 및 조성물
US20180237836A1 (en) * 2015-08-17 2018-08-23 The Regents Of The University Of California Microdroplet-Based Multiple Displacement Amplification (MDA) Methods and Related Compositions
US20190329209A1 (en) 2016-06-14 2019-10-31 Mission Bio, Inc. Methods and compositions for emulsification of solid supports in deformable beads
JP6785476B2 (ja) 2016-09-02 2020-11-18 国立大学法人 東京大学 検出装置及び細胞含有物体
JP6910788B2 (ja) 2016-12-06 2021-07-28 株式会社平和 遊技機
CN116622807A (zh) * 2023-05-26 2023-08-22 上海科技大学 一种单细胞全基因组测序文库的构建方法

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Publication number Publication date
WO2019216271A1 (fr) 2019-11-14
KR20210006418A (ko) 2021-01-18
US20210246495A1 (en) 2021-08-12
JPWO2019216271A1 (ja) 2020-10-22
EP3812460A1 (fr) 2021-04-28
JP6877804B2 (ja) 2021-05-26
SG11202011073RA (en) 2020-12-30
JP2021118717A (ja) 2021-08-12
AU2019265111A1 (en) 2021-01-07
EP3812460A4 (fr) 2021-08-11
CN112334578A (zh) 2021-02-05
US11142787B2 (en) 2021-10-12
TW202003829A (zh) 2020-01-16

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